水稻抗稻瘟病菌防卫反应的细胞学分析与防卫基因表达

对水稻类病变坏死突变体 lmm1　(LmmKaty)与原亲本及双单倍体YT14和YT16接种稻瘟病菌24 h内的抗病反应、细胞学和分子作用机理进行了研究。人工接种实验表明,与原亲本相比,该突变体对稻瘟病菌具有更强的抗性。突变体接种20~24 h后,自动荧光检测到细胞死亡,而对照则未检测到。表明该突变体通过细胞程序性死亡阻止稻瘟病菌的蔓延,从而提高了抗病性。Northern blotting结果发现,抗病品系YT14中的防卫相关基因、苯丙氨酸解氨酶基因和β 葡聚糖酶基因在人工接种稻瘟病菌6 h后开始表达,16~24 h后上述基因表达量快速增加;而病程相关蛋白基因 PR 1和几丁质酶基因在人工接种稻瘟病菌24 h后才开始表达。与YT14相比,感病品系中上述基因的表达明显延迟。表明在人工接种稻瘟病菌24 h内水稻可启动抗病防卫反应,从而对稻瘟病菌产生抗性。     

一个水稻类病变黄叶突变体的鉴定和精细定位

从15000多个水稻转基因株系中发现了一个类病变黄叶突变体.该突变体最显著的特征为叶片由下而上依次黄化,同时出现类似病原体感染的病斑.根据突变体表型,将该突变体命名为syll(spotted and yellow leayes 1).遗传学分析表明,该突变性状受1对隐性核基因控制.PCR检测和潮霉索抗性分析显示.该突变...     

水稻类病变突变体性状及分子机理研究进展

类病变突变体是植物在无显著逆境或损伤情况下以及未受到病原物危害时叶片上自然形成的类似病斑的一类突变体。水稻类病变突变体的产生常伴有抗病性的增强,其中涉及到防御相关基因的表达。相关研究主要集中于各种突变体的定位和克隆以及突变体的抗病性和抗病机制。综述了水稻类病变突变基因的性状及抗病性的研究进展,特别是对水稻类病变突变体形成原因、遗传特性、调控途径进行了梳理,并提出对水稻抗病育种的展望,以期为进一步分析类病变突变体的各种机制奠定理论基础,并为水稻育种提供参考。     

水稻发育障碍突变体a535di的细胞学与生理学研究

将水稻发育障碍突变体a535di与其F2正常显性植株做对比,突变体的花粉育性明显低于F2正常显性。扫描电镜观察发现突变体的花粉粒向内凹陷,且呈畸形,而F2正常显性的花粉粒则很饱满。突变体的超氧化物歧化酶（SOD）、过氧化物酶（POD）和过氧化氢酶（CAT）活性均低于F2正常显性植株,而出叶速度和叶绿素含量则不存在差异。     

一个水稻类病条纹斑突变体的鉴定和遗传定位

 粳稻品种嘉花1号经60Co γ射线辐射诱变后获得一个水稻类病斑突变体MR07, 暂命名为lms1。该突变体的病斑在全生育期均表现,属于扩散型类病斑突变体。生理和组织化学分析表明,该突变体在高温条件下（30℃）培养时只表现为白色条斑,低温条件下（20℃）培养时表现出细胞自主性死亡的坏死病斑。遗传分析表明,该突变体受1对隐性核基因控制。以lms1突变体与籼稻9311、培矮64S杂交的两个F2分离群体作为定位群体,利用SSR标记和Indel标记将该基因定位在第6染色体上,位于标记InDel1和MM0112-4之间,其物理距离为400 kb。     

水稻类病斑突变体lm8015-2的鉴定与基因的精细定位

[目的]为进一步解析水稻抗病分子机制,对类病斑突变体lm8015-2进行了鉴定.[方法]利用EMS诱变籼稻中恢8015,从其后代中鉴定出一个类病斑突变体lm8015-2.对突变体lm8015-2及其野生型的叶片进行超氧化物歧化酶活性、过氧化氢酶活性、过氧化物酶活性、过氧化氢含量、丙二醛含量、可溶性蛋白含量、光合色素含量...     

水稻类病斑突变体wy3的鉴定和基因定位

在粳稻品种武育粳3号栽培群体中获得一个类病斑突变体wy3。该突变体类病斑出现于苗期,分蘖期扩散至整张叶片,属于扩散型类病斑突变体。相比野生型,突变体wy3的株高明显降低,有效分蘖数减少,穗长、每穗粒数、结实率均显著降低。遮光处理表明,突变体wy3类病斑的产生受自然光诱导。台盼蓝染色结果表明,类病斑部位有大量的死亡细胞。突变体wy3的光合色素含量和净光合速率较野生型显著降低,SOD、POD、CAT活性和MDA含量均显著高于野生型。遗传分析表明突变体表型受单隐性核基因控制,采用BSA将该基因初步定位在第2染色体短臂端粒附近。采用F2群体中1099株类病斑单株将基因定位在标记W2-17和W2-18之间28kb的物理距离内。测序结果表明,突变体wy3中的LOC_Os02g02000编码区(CDS)第375位碱基C缺失,导致翻译提前终止,突变体中该候选基因为OsHPL3的一个新等位基因。     

水稻类病斑突变体lmm7的鉴定与基因定位

【目的】对水稻类病斑突变体的研究有助于解析其与植物生长和防御反应的关系。【方法】本研究在粳稻品系FI135胚培养过程中获得了1个类病斑突变体lmm7(lesion mimic mutant 7)。通过对其进行系统的表型鉴定、农艺性状考查、超微结构观察、生理学特性分析,阐明LMM7基因对植物生长的调控。通过病原菌抗性鉴定,明确lmm7对植物防御反应的影响。利用9311B与突变体lmm7杂交所得F₂群体对该突变体进行了遗传分析和基因精细定位。【结果】该突变体苗期表型正常,分蘖初期,植株基部叶片从叶尖开始不断出现褐色斑点,并向整株扩散,且斑点数目随植株生长不断增加。与野生型相比,突变体的株高、穗长、有效穗数、每穗粒数、结实率及剑叶长宽都显著降低,但籽粒性状和抽穗期没有显著性差异。遮光处理表明,突变体lmm7的表型受到光照诱导,抽穗期突变体lmm7叶肉细胞严重失绿,光合色素含量显著降低。组织化学分析表明,突变体病斑处的H₂O₂含量显著升高。透射电镜观察结果表明,突变体lmm7叶肉细胞的叶绿体数目减少,叶绿体类囊体片层结构严重受损,细胞器肿胀解体,并出现大量嗜锇小体,同时病斑内部和周围区域积累了大量的ROS。抗性鉴定结果显示突变体lmm7稻瘟病抗性水平显著高于野生型。遗传分析表明lmm7的突变表型受单个隐性基因控制。利用图位克隆的方法,目的基因被定位于水稻第7染色体短臂两InDel标记7B35和7B43之间,区间范围约260 kb。测序结果表明该区间内候选基因LOC_Os07g0203700第2891位碱基T发生了单碱基缺失,导致后续移码突变及翻译提前终止。【结论】lmm7与spl5互为等位基因,其突变抑制了植株的生长,同时增强了对稻瘟病的抗性。     

大豆雄性不育突变体NJ89—1的遗传学与细胞学鉴定

花粉发芽实验ＮＪ８９－１雄性不育度高达９９．２５％以上且稳定，人工授粉试验ＮＪ８９－１的雌性育性正常，说明ＮＵ８９－１是一个雄性不育雌性可育突变体。不育株自然粉后代中育性分离结果表明ＮＪ８９－１雄性不育性受单隐性核基因控制。     

水稻雄性不育突变体TP79的遗传分析及细胞学研究

TP79是一个源于水稻品种台北309的自然突变的不育突变体,对该突变体进行遗传分析和细胞学研究,结果显示,TP79小花雌蕊正常,雄蕊6枚,花丝细长,花药干瘪,花粉以染败类型为主;利用TP79与台北309杂交进行遗传分析,结果表明,TP79是单隐形核基因控制的雄性不育突变体;组织切片观察发现,TP79在小孢子形成前期出现异常,绒毡层不能正常降解,小孢子的发育畸形,在最终花粉成熟期,绒毡层仍呈浓缩状,形成的花粉干瘪,无活性。     

Characterization and Proteomic Analysis of Novel Rice Lesion Mimic Mutant with Enhanced Disease Resistance

Lesion mimic mutants(LMMs) are plants that spontaneously form lesions without pathogen infection or external stimulus and exhibit resistance to pathogens. Here, a rice LMM was created by ethyl methane sulfonate mutagenesis, named as hpil(hydrogen peroxide induced lesion). Diaminobenzidine and trypan blue staining showed that large amounts of H_2O_2 were produced and cell death was occurred at and around the parts of lesion mimic in the rice leaves. The phenotype of hpil is controlled by a single recessive gene, localized at a 2 Mb interval on chromosome 2. The data suggested that hpil is a novel LMM with enhanced bacterial and fungal disease resistance, and multiple pathogenesis-related proteins(PRs) were up-regulated. The proteomes of leaves at three positions(different degrees of lesion mimic severity) were characterized in hpil compared with its wild type plant. Differentially expressed proteins were detected by two dimensional difference gel electrophoresis and 274 proteins were identified by MALDI TOF/TOF~(TM). These proteins were related to metabolic process, cellular process and response to stimulus, with mostly down-regulated in hpil leaves. Many of these proteins were related to the Calvin cycle, photosynthetic electron transport chain, glycolysis/gluconeogenesis and phosphonates pathways. Some resistance-related proteins including 14-3-3 proteins, OsPR10 and antioxidases such as peroxidase, superoxide dismutase and ascorbate peroxidase were up-regulated in leaves with lesion mimic. These results provide the foundation for cloning of the target gene and shed light on the mechanism involved in autoimmunity of rice.     

Identification and Genetic Mapping of a Lesion Mimic Mutant in Rice

A lesion mimic stripe mutant,designated as lms1(lesion mimic stripe 1),was obtained from the M2 progeny of a 60Co γ-radiation treated japonica rice variety Jiahua 1.The lms1 mutant displayed propagation type lesions across the whole growth and developmental stages.Physiology and histochemistry analysis showed that the mutant exhibited a phenotype of white stripe when grown under high temperature(30 ℃),and the lesion mimic caused by programmed cell death under low temperature(20 ℃).The genetic analysis indicated that this lesion-mimic phenotype is controlled by a single locus recessive nuclear gene.Furthermore,by using simple sequence repeat markers and an F2 segregating population derived from two crosses of lms1 × 93-11 and lms1 × Pei'ai 64S,the lms1 gene was mapped between markers Indel1 and MM0112-4 with a physical distance of 400 kb on chromosome 6 in rice.     

Morphological Structure and Genetic Mapping of New Leaf-Color Mutant Gene in Rice (Oryza sativa)

Leaf-color mutations are a widely-observed class of mutations, playing an important role in the study of chlorophyll biosynthesis and plant chloroplast structure, function, genetics and development. A naturally-occurring leaf-color rice mutant, Baihuaidao 7, was analyzed. Mutant plants typically exhibited a green-white-green leaf-color progression, but this phenotype was only expressed in the presence of a stress signal induced by mechanical scarification such as transplantation. Prior to the appearance of white leaves, mutant plant growth, leaf color, chlorophyll content, and chloroplast ultrastructure appeared to be identical to those of the wild type. After the changeover to white leaf color, an examination of the mutated leaves revealed a decrease in total chlorophyll, chlorophyll a, chlorophyll b, and carotenoid content, a reduction in the number of chloroplast grana lamella and grana, and a gradual degradation of the thylakoid lamellas. At maturity, the mutant plant was etiolated and dwarfed compared with wild-type plants. Genetic analysis indicated that the leaf mutant character is controlled by a recessive nuclear gene. Genetic mapping of the mutant gene was performed using an F2 population derived from a Baihuaidao 7 × Jiangxi 1587 cross. The mutant gene was mapped to rice chromosome 11, positioned between InDel markers L59.2-7 and L64.8-11, which are separated by approximately 740.5 kb. The mutant gene is believed to be a new leaf-color mutant gene in rice, and is tentatively designated as gwgl.